Contamination by Microplastics in Free-Living White-Eared Opossums (Didelphis Albiventris) Resident in Campo Grande, Mato Grosso Do Sul – Brazil

Contamination by Microplastics in Free-Living White-Eared Opossums (Didelphis Albiventris) Resident in Campo Grande, Mato Grosso Do Sul – Brazil | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Contamination by Microplastics in Free-Living White-Eared Opossums (Didelphis Albiventris) Resident in Campo Grande, Mato Grosso Do Sul – Brazil Fernanda Mayara Gauto Melo, Heitor Miraglia Herrera, Amanda Costa Rodrigues, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7284106/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 22 Oct, 2025 Read the published version in Environmental Monitoring and Assessment → Version 1 posted 11 You are reading this latest preprint version Abstract This study aimed to investigate the presence of microplastics in the gastrointestinal tract and lungs of white-eared opossums ( Didelphis albiventris ), as well as their effects on these organs and body condition of sampled animals. We captured 22 opossums from April to December 2023 in six forest fragments of Campo Grande, capital of Mato Grosso do Sul state. The estimate of body condition was performed using measurements of weight and head-body length for males and females. Stomach, ileum, and lung fragments were collected during necropsies for histopathological study and detection of microplastics by alkaline digestion with KOH. Digested samples were vacuum filtered using filter paper and dried in an oven. The dried filters were examined under an optical microscope. A total of 270 microplastics were detected in the 22 animals examined. The ileum presented the highest number of microplastics (n = 141), fibers were the most common type (n = 185), and blue was the predominant color (n = 140). Regarding chemical composition, the most frequently detected microplastics were polyvinyl chloride, polyethylene terephthalate, and acrylonitrile butadiene styrene. Although all D. albiventris presented microplastics in their evaluable tissues, no effects on body condition or tissue damage were observed. Since D. albiventris are common animals in Campo Grande and are contaminated by microplastics, these animals can act as sentinels of environmental condition. Bioindicator Body condition Emerging contaminant Histopathology Polyethylene Wild mammal Figures Figure 1 Figure 2 Figure 3 Introduction As the global population grows exponentially, a large number of pollutants threatens the health of humans and animals, such as polymers, that are widely found in the terrestrial (Beriot et al. 2021 ; Huerta Lwanga et al. 2016 ) and aquatic environment (Allen et al. 2022 ; Cózar et al. 2014 ; Kibria et al. 2023 ; Martí et al. 2020 ). Indeed, because of the chemical polymers used to manufacture plastic originate from non-biodegradable sources (Andreeßen and Steinbüchel 2019 ), micrometer-sized particles known as microplastics (MPs) (Kühnel et al. 2023 ) remain in the environment and are easily absorbed by living organisms, such as humans, plants and fish (S. Singh and Tiwari 2025 ). Plastics can be classified as thermosets and thermoplastics. Thermosets include phenol-formaldehyde resin (bakelite), epoxy (araldite), and polyurethane (PUR), while thermoplastics include polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyethylene terephthalate (PET), polystyrene (PS), high-density polyethylene (HDPE), low-density polyethylene (LDPE), polycarbonate (PC), polyamide (PA), polymethyl methacrylate (PMMA), acrylonitrile butadiene styrene (ABS), acrylonitrile styrene acrylate (ASA), styrene-acrylonitrile copolymer (SAN), and thermoplastic elastomers (TPE) (Plastic Europe n.d.). Thermoplastics are the most prevalent in the environment, as they are the most widely used in the market due to their extensive use in disposable packaging (Cruz and Zanin 2003 ). As a polymer, plastic degradation processes drastically reduce its molecular mass (Cózar et al. 2014 ), leading to fissure formation and increased wettability and crystallinity (Waldman and Rillig 2020 ). These processes can be physicochemical, biological, and mainly mechanical, due to erosion caused by wind, waves, and solar radiation (photolysis) (Moore 2008 ), generating microplastics (MPs) that are omnipresent and persistent anthropogenic contaminants found in urban environments (Dris et al. 2018 ). Furthermore, MPs can cause various pathological alterations due to tissue abrasion by plastic debris, thereby triggering inflammatory responses, insufficiency and reduced energy levels, as well as lower growth rates (Huerta Lwanga et al. 2016 ; Y. Song et al. 2019 ; Welden and Cowie 2016 ). Various techniques have been employed to identify MPs based on their chemical composition and specific functional groups (Bolan et al. 2020 ). These analyses are conducted using advanced equipment such as stereoscopes, scanning electron microscopes, fluorescence microscopes, Raman/FTIR microscopes, and matrix-assisted laser desorption/ionization (Huppertsberg and Knepper 2018 ; Mukotaka et al. 2021 ; Siegel et al. 2021 ; Y. K. Song et al. 2015 ). Fourier-transform infrared (FT-IR) imaging spectroscopy has remained the most widely applied method in microplastic research (Renner et al. 2018 ). This method has proven effective for detecting, measuring, and analyzing MP contamination in aquatic and terrestrial communities (Horton et al. 2021 ; Napper et al. 2020 ; Peeken et al. 2018 ; Weisser et al. 2021 ). Furthermore, the use of histopathological examinations is necessary to localize microplastics within tissues enhancing the knowledge of its effect in the organism, contrasting with extraction-based approaches that only provide a general overview of particle presence in an organ or tissue (Prata et al. 2022 ). This technique also allows observation of tissue alterations caused by the presence of microplastics in different organs and systems (Lee et al. 2025 ). The white-eared opossum ( Didelphis albiventris ) is a common marsupial in the municipality of Campo Grande, Mato Grosso do Sul (MS), Brazil (Cáceres 2002 ; Sanches et al. 2022 ). It is considered as synantropic animal because they can be favored by human presence, Didelphis spp. can be found in urban parks and gardens, vacant lots, in peri-domicile and in human and animal dwellings. Furthermore, as Didelphis spp. are omnivorous animals, they feed on a wide variety of items, including fruits, insects, eggs, and small vertebrates (Cáceres 2002 ). Contamination by MPs can occur through various routes, such as ingestion, inhalation, dermal contact, transplacental transfer, and predation of contaminated animals. For many mammal species, ingestion is described as the primary contamination pathway (Meaza et al. 2021 ). Each contamination route carries specific risks and potential health consequences for the animals and humans (Rahman et al. 2021 ). In this context, the present study aimed to investigate the presence of MPs in the gastrointestinal tract and lungs of D. albiventris sampled in an urban area of Campo Grande, MS. Furthermore, we evaluated the effects of MPs on those tissues and Body Condition (BC) of sampled animals, providing insights into the influence of MPs in the health of this synantropic species in anthropized areas of Mato Grosso do Sul State, Brazil. Materials and Methods Study Area and Data Collection This study was carried out from April to December 2023 in six forest fragments in the municipality of Campo Grande, MS [Reserva do Surucuá (20°24'47"S 54°36'55"W), CEA Polonês (20°26'49"S 54°34'48"W), Clube Carandá (20°26'21"S 54°34'27"W), Chácara Coqueiral (20°29'34"S 54°34'53"W), CEA Imbirussu (20°26'57"S 54°41'48"W), and CEA Anhanduí (20°30'22"S 54°38'37"W)]. All fragments were sampled at least three times. We used 20 wired live-trap baited with bacon and revised twice a day. Traps remained set for five consecutive nights in each forest fragment, it were installed on the ground and placed approximately 10 meters apart. Captured females were previously evaluated for pregnancy and released if there were cubs attached in their pouch. Individuals were anesthetized intramuscularly with ketamine (20 mg/kg) and xylazine (2 mg/kg). Subsequently, individual and biometric data were collected, including head-body length, body-tail length, weight and sex. Afterwards, animals were euthanized by intravenous administration of potassium chloride (75–150 mg/kg), following the UNIFESP euthanasia protocol. During necropsy, lung, ileum, and stomach (preserving contents) were collected. Samples contemplated for MPs examination were weighed in fragments ranging from 0.13 g to 7.49 g (lung), 0.37 g to 5.76 g (ileum), and 0.5 g to 12.5 g (stomach), wrapped in aluminum foil, and frozen at − 20°C until processing. Fragments of the same organs measuring approximately two centimeters in thickness, length, and width were fixed in 10% formalin for histopathological examination. All procedures carried out during this research complied with the Biodiversity Authorization and Information System (SISBIO) protocol numbers 70946-5 and 89586-1. Sample handling and processing followed the guidelines established by the Ethics Committee on Animal Use (CEUA) of Dom Bosco Catholic University, protocol number 013/2020. Sample Processing Stomach, ileum, and lung samples were thawed at room temperature. We used two control filters for each group of organ fragments: the first to monitor air and environmental contamination, and the second was subjected to filtration with 10% KOH solution and Milli-Q water to ensure solution purity. For MPs assessment, biological samples were digested using a 10% potassium hydroxide (KOH) solution (Kühn et al. 2017 ). Samples were rinsed with Milli-Q water to remove residues, weighted on a semi-analytical balance (Marte Ad500 510 g × 0.001 g), and placed in glass beakers with 10% KOH solution at a ratio of three times the biological mass (3:1) (Dehaut et al. 2016 ). Beakers were covered with aluminum foil and agitated twice daily at room temperature until complete digestion. After digestion, the solution was vacuum filtered using a 47 mm glass fiber microfilter GF-1 (Macherey Nagel) with a pore size of 0.7 µm. Each microfilter was placed individually in glass Petri dishes and dried in an oven for 1 h at 70°C. Filters were examined using a Carl Zeiss optical microscope (MOC), model Axio Scope A1, and images were captured with Zen software (software version to be verified). Tissue images were taken with a color camera (Axiocam 503) attached to the MOC. The materials found were counted and classified following the protocol used by (Clere et al. 2022 ). Fibers were measured for length and width, while fragments were measured by area. After filter analysis, filters were washed with Milli-Q water in beakers and pooling samples of all animals were separated by organ. Filters were then oven-dried at 70°C until completely dry. Samples were sent to the Chemistry Institute at the Federal University of Mato Grosso do Sul (UFMS) for FT-IR spectroscopic analysis. Fourier Transform Infrared Spectroscopy Spectral data in the mid-infrared region were obtained using a Perkin Elmer Frontier FT/IR spectrometer with Fourier transform. Analyses were acquired over the spectral window from 500 to 4000 cm⁻¹. Samples were prepared by dispersion in KBr pellets. The obtained spectra were compared with those from the reference library created by (Shimadzu n.d.). Only spectra with a match of 70% or higher to the standard database were considered reliable and recorded as verified MPs (Zhao et al. 2018 ) Histopathological Examination For histopathological examination, fragments of lung, ileum, and stomach were collected. Tissues were sectioned into fragments measuring 3 mm to 5 mm, placed in cassettes, and sent to the Technical Histopathology Center of Curitiba - PR, Brazil, for slide preparation and Hematoxylin and Eosin (H&E) staining. Histopathological slides were analyzed in the laboratory of Dom Bosco Catholic University using a Carl Zeiss optical light microscope (MOC), model Axio Scope A1, and images were captured with the aid of Zen software. Tissue images were photographed using an Axiocam 503 color camera attached to the MOC. Statistical Analysis A set of models was constructed to evaluate the effects of MPs size, found in organs analyzed individually and combined, on the body condition (BC) of the collected D. albiventris . BC was used as a health assessment parameter and calculated by means of the standardized residuals from a linear regression between body weight and head-body length, computed separately for males and females. Additionally, a null model without explanatory variables was created. Generalized linear models (GLMs) were ranked using an approach based on Akaike’s Information Criterion corrected for small sample sizes (AICc) (Akaike 1974 ), considering all models with ΔAICc ≤ 2 as plausible (Burnham and Anderson 2004 ). The ‘AICcmodavg’ package version 2.3–1 in R 3.5.0 (R Core Team 2021 ) was used for model ranking. Results and Discussion Our results show that terrestrial wildlife found in urban areas can be found contaminated by MPs since we found by tissue digestion a total of 270 MPs in the gastrointestinal tract and lungs of the all 22 white-eared opossums captured in the municipality of Campo Grande-MS. In fact, until now, contamination by MPs in terrestrial animals was reported only in domestic animals as sheep (Beriot et al. 2021 ), dogs and cats (Zhang et al. 2019 ), and chicken (Lwanga et al. 2016 ), as well as in earthworms (Lwanga et al. 2018 ). Most reports concern MPs contamination refer to marine animals such as bivalves (Li et al. 2015 ), seals (Perez-Venegas et al. 2018 ), commercial fish (Neves et al. 2015 ), pelagic and demersal fish (Neves et al. 2015 ), sea anemones (Morais et al. 2020 ), rays (Pegado et al. 2021 ), and shrimp (Hossain et al. 2020 ). Furthermore, since it has been suggested that the accumulation of MPs in the gastrointestinal tract of raptor birds may be due to a combination of direct ingestion of plastics and/or indirect consumption through trophic transfer (Carlin et al. 2020 ). Didelphis albiventris can also present both routes of contamination by MPs because they feed on small vertebrates and invertebrates. The high number of MPs found in D. albiventris found in this study may be associated to the omnivorous feeding behavior of this synanthropic species, in accordance to recorded in other vertebrates’ animals with omnivorous diets, which tend to be broader and less selective (Mazariegos Ortíz et al. 2021 ; Mizraji et al. 2017 ). The use of plastic debris from human consumption, such as plastic bags, has been already reported in nests of D. albiventris (Blettler and Mitchell 2021 ). The ileum was the tissue with the highest quantity of microplastics, with 141 MPs recorded (52%), followed by the stomach with 86 MPs (32%) and the lungs with 43 MPs (16%). The high number of MPs in the gastrointestinal tract of D. albiventris in this study may indicate that ingestion was the most common contamination route. Indeed, for various mammal species, ingestion is considered the primary exposure pathway (Meaza et al. 2021 ), with MPs most frequently detected in the digestive tract, especially in the ileum probably due to the presence of microfold cells located in the epithelium covering mucosa-associated lymphoid tissues, such as the Peyer's patches located in the ileum, that actively transport luminal antigens to the underlying lymphoid follicles to initiate an immune response (Arumugasaamy et al. 2018 ; Smith et al. 1995 ). Fibers were the most common type of MPs found (68.5%), followed by fragments (30.4%), foam (0.7%), and film (0.4%). Fibers was found in 79.4% in ileum sampled, 62.8% lungs, and 53.5% stomach. Studies report a higher prevalence of MPs fibers in the intestinal tract of cows and sheep in Iran (Bahrani et al. 2023 ), intestinal tract of ducks in Indonesia (Susanti et al. 2021 ), as well as in the gastrointestinal tract of terrestrial birds in Florida, USA (Carlin et al. 2020 ) and Shanghai, China (Zhao et al. 2016 ). Fibers have high bioaccumulation potential and are among the most common types observed in both marine environments (Browne et al. 2011 ) and soil (R. P. Singh et al. 2020 ; Zhou et al. 2020 ). A study on the mussel Mytilus galloprovincialis showed that fibers persisted in the organism longer than fragments and spheres (Park et al. 2024 ). Research on zebrafish demonstrated that fiber accumulation in the intestine was greater compared to fragments and spheres, causing more severe intestinal toxicity (Qiao et al. 2019 ). Additionally, fibers have been reported in high amounts in lung tissue of pigs and humans with lung cancer (Pauly et al. 1998 ; Li et al. 2023 ). MP fibers were found in various colors (Fig. 1 ), blue was the most abundant one comprising 52% of the particles, appearing most frequently in the lungs and ileum. Transparent MPs were the second most abundant polymer, accounting for 16%, followed by black (14%), amber (7.4%), red (6%), orange (1.8%), purple, and gray (0.7%). Blue fibers have been frequently reported (Araujo Dutra and Camargo Maia 2023 ; Martí et al. 2020 ) probably because blue plastics tend to degrade faster under sunlight, as they do not absorb UV light effectively, increasing the fragmentation rate and consequently the amount in the environment (Martí et al. 2020 ; Yang et al. 2022 ). The second most common morphological type of MPs was fragments. Like fibers, they were also detected in the lungs, ileum, and stomach. The predominant colors were blue, black, amber, purple, multicolored, transparent, and gray (Fig. 2 ). The presence of amber coloration in MPs may be related to the photoaging of plastics, which gradually alters their color (discoloration) during prolonged solar exposure (Martí et al. 2020 ), considering that detection of amber-colored MPs in the gastrointestinal tract of animals is rarely reported. Plastic color significantly affects the absorption of sunlight, with plastics of different colors absorbing light at different wavelengths (Zhao et al. 2022 ). Regarding FT-IR analyses, three types of polymers were identified: polyethylene terephthalate (PET), acrylonitrile butadiene styrene (ABS), and polyvinyl chloride (PVC). These polymers are classified as thermoplastics, the most commonly used in disposable packaging industries (Plastic Europe n.d.). We observed that all samples contained PVC (Supplementary Material – Figures S1 , S2 and S3). While PET was found in the ileum and lungs (Supplementary Material – Figures S2 and S3), ABS was detected only in lungs (Supplementary Material – Figure S3). The presence of PVC and PET in the gastrointestinal tract and lungs of D. albiventris can be attributed to the high abundance of these polymers in the environment (Akdogan and Guven 2019 ), as they are among the most commonly found MPs in both soil (Yang et al. 2022 ) and air (Yao et al. 2022). Indeed, D. albiventris are frequently found in trash bins in Campo Grande, potentially ingesting PVC and PET from plastic commonly used packaging (Wang et al. 2021 ). The presence of ABS in animals has not been previously reported in the literature. Furthermore, reports of this polymer in sediments and effluents, are also scarce (Belesov et al. 2024 ; Sarti et al. 2024 ). Since we found ABS only in the lungs, it seems that the airborne contamination route would be the most likely. We highlight that MPs have been identified in a wide range of species (Table 1 ), from urban dogs and cats to dolphins, birds of prey, terrestrial mammals, and commercial fish, covering different geographic regions and habitats. The increasing detection of microplastics in various tissues and organs of animals reinforces the urgency of studies focused on pollution caused by these emerging contaminants. The most common polymers include PET, PE, PP, and PVC, with a predominance of synthetic fibers and fragments smaller than 1 mm. The presence of these materials in vital organs such as lungs, liver, gastrointestinal tract, and reproductive tissues raises concerns about their toxic, bioaccumulative effects and potential consequences on animal health and, by extension, human health, given the ecological interconnectedness. These data emphasize that the ubiquity of MPs in the environment is already directly reflected in biota, making it essential to deepen investigations into exposure mechanisms, accumulation, and toxicity, as well as to strengthen public policies and mitigation actions against this form of pollution. Table 1 Studies on the detection of microplastics in tissues and organs of different animal species across various countries. Animals Location Polymer Types Organs Techniques Uses Remarks Authors White-eared opossum Mato Grosso do Sul, Brazil PET, PVC and ABS Stomach, ileum, lung FT-IR 100% showed MPs in at least one organ, highlighting their role as sentinels of contamination, potentially exposed via ingestion and inhalation. This study Urban dogs and cats Porto, Portugal PET, PP Lung, ileum, liver, kidneys, blood clots Micro-Raman spectroscopy Microplastics were found in 100% of the internal tissues analyzed, predominantly synthetic fibers, indicating high environmental exposure and potential health risks to urban animals. Prata et al., 2022 Dogs and humans New México, USA PE, PVC, PET Testicular tissues Py-GC/MS Presence of MPs in the testes, decreased sperm count, and reduced testicular and epididymal weight, suggesting potential impacts on male reproductive health Hu et al., 2024 Terrestrial birds of prey Florida, USA PET, PE GI tract µ-FTIR Generalized accumulation of MPs in the gastrointestinal tract, with 100% of samples contaminated, indicating wide environmental exposure and potential health risks. Carlin et al., 2020 Wild terrestrial animals Thailand PE, PET, PP, PVDC, PES, PU Carcasses (organs not specified) FT-IR MPs found in 71% of fecal samples, demonstrating that plastic contamination already widely affects the terrestrial fauna in the region. Teampanpong & Duengkae, 2024 Dolphins ( Tursiops truncatus ) Florida, USA Polyester Gastrointestinal fluid FT-IR Presence of MPs in muscular and gastrointestinal tissues of fish from Sarasota Bay, the dolphins' main food source, suggesting risk of indirect exposure. Hart et al., 2022 Commercial fish Xisha, China PE, PET, PVC, PP, PVA Gills and GI tract FT-IR MPs detected in 61.7% of reef fish, primarily in the gastrointestinal tract, with a predominance of polyamide and PET fibers. Huang et al., 2023 Wild coastal mammals, birds, and fish Bergen, Norway PET, PVC, PC, PP, PE, PS, PMMA, PA Stomach, intestinal wall, liver, muscle Py-GC/MS MPs present in 62% of analyzed tissues, with higher concentrations in the liver and a predominance of PVC, PS, and PET, indicating potential bioaccumulation in internal organs. Haave et al., 2021 ) Coyote, Virginia opossum, white-nosed coati, white-tailed deer, Mexican and lesser anteater Yucatán, Mexico Elastomers, PS, HDPE, cellulosic polymers, PU Feces FT-IR MPs detected in 100% of terrestrial vertebrate fecal samples, predominantly fibers < 1 mm composed of polyamide and PET, revealing widespread environmental contamination. Mendoza-Arroyo et al., 2024 Terrestrial mammals United Kingdom Polyester, PE, PNB, and biodegradable plastics Feces µ-FTIR The study revealed that 16.5% of the terrestrial mammals ingested MPs, highlighting extensive exposure to environmental pollution. Thrift et al., 2022 Note. PET = polyethylene terephthalate; PE = polyethylene; PP = polypropylene; PVC = polyvinyl chloride; PS = polystyrene; PU = polyurethane; PVA = polyvinyl acetate; PMMA = polymethyl methacrylate; PNB = polybutylene; ABS = acrylonitrile butadiene styrene. FT-IR = Fourier-transform infrared spectroscopy; µ-FTIR = micro-Fourier-transform infrared spectroscopy; Py-GC/MS = pyrolysis coupled with gas chromatography and mass spectrometry. GI tract = gastrointestinal tract. The histopathological examination of sampled D. albiventris revealed the presence of few MPs in only intestinal interstitium and the lumen of the stomach, amid the contents, with no inflammatory reaction observed in the organs (Fig. 3 ). These findings are in agreement with Haave et al. ( 2021 ) that did not find MPs or tissue reaction by histopathological examination in fish, seabirds, terrestrial and marine mammals from a coastal area of Norway. The number of MPs observed in the histopathology of the gastrointestinal tract of D. albiventris , in this study, seems insufficient to cause intestinal blockage or a feeling of fullness. The pulmonary parenchyma of D. albiventris did not showed neither MPs nor alterations. Most studies evaluating histopathological effects of MPs are experimental, conducted in laboratory settings (Ali et al. 2023 ; Hamed et al. 2021 ), aimed to induce tissue damage (Hoseini et al. 2022 ; Saleh et al. 2025 ; Sayed et al. 2022 ). These studies use high concentrations of commercial MPs, that are not commonly found in the environment (Allen et al. 2022 ; Sun et al. 2021 ). The contamination of MPs in D. albiventris does not seem to threaten health of sampled animals since our statistical analyses did not correlate the presence of MPs in tissues with BC (Supplementary Material, Table S1 ). This corroborates with de Vries et al. ( 2020 ) in cod and hake contaminated with MPs, which hypothesized that, especially in larger individuals, MPs are not retained, thus not affecting BC, unlike what occurs in small fish species (Critchell and Hoogenboom 2018 ). Conclusion All individuals of D. albiventris were found contaminated by MPs, without effects on health of sampled animals. Moreover, this synanthropic species may be used as sentinels of environmental contamination in anthropized areas. Declarations Ethical Approval The research was approved by the Animal Use Ethics Committee of Dom Bosco Catholic University (protocol 013/2020) and the Biodiversity Authorization and Information System (SISBIO) (protocol 70946-5 and 89586-1). The authors declare no relevant financial or non-financial interests. Conflict of Interest Statement The author(s) declared no potential conflicts of interest regarding the research, authorship, and/or publication of this article. Authors’ Ethical Responsibilities All authors have read, understood, and complied, as applicable, with the statement on "Authors' Ethical Responsibilities," as found in the Instructions for Authors. Funding Declaration This work was supported by National Council for Scientific and Technological Development (CNPq) and Coordination of Improvement of Higher Education Personnel (CAPES). ACR received a fellowship from CNPq (163253/2022-06). FMS received a fellowship from Funding Authority for Studies and Projects (FINEP) (01.24.0114.00) and CNPq (Productivity Scholarship 302176/2025-0), PSC received a fellowship from Ministry of Science, Technology and Innovation (MCTI) (352739/2025-8) and HMH is researcher from CNPq (Productivity Scholarship 311769/2023-3). Author Contribution F.M.G.M., A.C.R., G.B.A., P.S.C., and H.M.H. contributed to the conceptualization of the study. F.M.G.M., P.S.C., A.C.R., O.F.J., and W.A.G.N. were responsible for the methodology. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7284106","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":502723679,"identity":"cf50cf71-5f9f-473f-941e-614e64edc367","order_by":0,"name":"Fernanda Mayara Gauto Melo","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Fernanda","middleName":"Mayara Gauto","lastName":"Melo","suffix":""},{"id":502723680,"identity":"47fa44df-e3cf-4dc0-ad2a-0791e832851a","order_by":1,"name":"Heitor Miraglia Herrera","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Heitor","middleName":"Miraglia","lastName":"Herrera","suffix":""},{"id":502723681,"identity":"e631aa5f-b2b4-4546-b205-78165acdbcdd","order_by":2,"name":"Amanda Costa Rodrigues","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Amanda","middleName":"Costa","lastName":"Rodrigues","suffix":""},{"id":502723682,"identity":"673dcaf5-d63f-4ab8-8696-04faca947074","order_by":3,"name":"Oscar Fernandes Junior","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Oscar","middleName":"Fernandes","lastName":"Junior","suffix":""},{"id":502723683,"identity":"e9df39de-d238-4b95-8b0e-ac8743e545a9","order_by":4,"name":"Filipe Martins Santos","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Filipe","middleName":"Martins","lastName":"Santos","suffix":""},{"id":502723684,"identity":"2c6e6ec4-e35a-4f73-8c06-376ac6207421","order_by":5,"name":"Taynara Gabrielly Santos Pinto","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Taynara","middleName":"Gabrielly Santos","lastName":"Pinto","suffix":""},{"id":502723685,"identity":"378fdd44-767a-4426-8a0f-7a6a8ff44018","order_by":6,"name":"Wesley Arruda Gimenes Nantes","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Wesley","middleName":"Arruda Gimenes","lastName":"Nantes","suffix":""},{"id":502723686,"identity":"46a933ad-0cb0-4497-b37d-5769b7458853","order_by":7,"name":"Nayara Yoshie Sano","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Nayara","middleName":"Yoshie","lastName":"Sano","suffix":""},{"id":502723687,"identity":"e1cb1beb-0b15-4f72-a113-42fb5e7af908","order_by":8,"name":"Priscila Sabioni Cavalheri","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABE0lEQVRIie2RMUvDUBDHLzxIliuuJyntV7gQMA0ofpUEwalD3TrVQCEuQdd8DKfMCW9w6QdwcJFAJodKQTJ08CwVRJK4OrzfdMefH3f3HoDB8C+xEgCeHZtFCaBUCUupeVihY8Nfih3BZlA58FMB5EElcPV62y5oCo5uqOWXSXCHH1TNVxKVnUp4H6enGZOX4PWZFI0/1qOCqkJLFHUqvLFSJbdYCYHPyDrO1ahw34tSou7FRFnvRLlMyNl5e9a3ucJGpqyGlMQVJU4IvVqmRKTQFkX1KmFmHW65SnF+U49Ze7my/ZncgmHW82Lo1Nt2f37x4Dw9Vm9LPaUT/fosi00C7Fnsu7B/Jz3Cn19sMBgMBuET2KRWsY+RKM0AAAAASUVORK5CYII=","orcid":"","institution":"Federal University of Mato Grosso do Sul","correspondingAuthor":true,"prefix":"","firstName":"Priscila","middleName":"Sabioni","lastName":"Cavalheri","suffix":""},{"id":502723688,"identity":"677ea2c3-b253-473a-ba74-536fec809a4d","order_by":9,"name":"Gisele Braziliano Andrade","email":"","orcid":"","institution":"Dom Bosco Catholic University","correspondingAuthor":false,"prefix":"","firstName":"Gisele","middleName":"Braziliano","lastName":"Andrade","suffix":""}],"badges":[],"createdAt":"2025-08-03 14:53:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7284106/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7284106/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10661-025-14682-3","type":"published","date":"2025-10-22T16:17:10+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":89668289,"identity":"26ff7da0-72c2-4b82-8679-89c6b37901d0","added_by":"auto","created_at":"2025-08-22 12:24:03","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":130660,"visible":true,"origin":"","legend":"\u003cp\u003ePhotomicrographs of the gastrointestinal tract and lung of \u003cem\u003eDidelphis albiventris \u003c/em\u003esampled at Campo Grande, Mato Grosso do Sul State, Brazil, showing microplastic fibers of different colors. Stomach: red fiber (a); Ileum: blue fiber (b), black fiber (c) and transparent fiber (d); Lung: royal blue fiber (e) and red fiber (f).\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7284106/v1/bc70abdf952b59421096cf90.jpeg"},{"id":89668297,"identity":"e16d9f4e-4cc9-424c-9828-e44c1b6bded5","added_by":"auto","created_at":"2025-08-22 12:24:04","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":3102722,"visible":true,"origin":"","legend":"\u003cp\u003ePhotomicrographs of the gastrointestinal tract and lung of \u003cem\u003eDidelphis albiventris\u003c/em\u003e sampled at Campo Grande, capital of Mato Grosso do Sul, Brazil, showing different shapes and colors of microplastic fragments. Stomach: transparent fragment (a) and gray fragment (b); Ileum: black fragment (c), blue fragment (d), black fragment (e) and blue fragment with yellow dots (f); Lung: purple fragment (g), blue fragment (h) and amber fragment (i).\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7284106/v1/b3d4e25a63b9d5161753e2c7.png"},{"id":89668279,"identity":"b9f15985-ec0c-4a95-81f5-5d0ac0300b1d","added_by":"auto","created_at":"2025-08-22 12:24:02","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":112819,"visible":true,"origin":"","legend":"\u003cp\u003ePhotomicrograph showing the presence of MPs in the gastrointestinal tract of \u003cem\u003eDidelphis albiventris \u003c/em\u003esampled at Campo Grande, capital of Mato Grosso do Sul State, Brazil. (a) Intestine displaying a black-colored fragment of microplastic adjacent to the intestinal gland (arrow); (b) Microplastic fragments in blue and amber coloration. Hematoxylin and eosin staining.\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7284106/v1/ff054f28fd796f53d5e53ed2.jpeg"},{"id":94490401,"identity":"2d3f703f-8e32-49aa-93b3-a0746121f2b3","added_by":"auto","created_at":"2025-10-27 17:09:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3939449,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7284106/v1/82939358-2660-4730-9d74-216c0bde0a79.pdf"},{"id":89668282,"identity":"7b947c7a-ccf2-4557-b242-8260d6469c99","added_by":"auto","created_at":"2025-08-22 12:24:02","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":78925,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-7284106/v1/2cb74e26a8c0c98347d28441.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eContamination by Microplastics in Free-Living White-Eared Opossums (Didelphis Albiventris) Resident in Campo Grande, Mato Grosso Do Sul – Brazil\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAs the global population grows exponentially, a large number of pollutants threatens the health of humans and animals, such as polymers, that are widely found in the terrestrial (Beriot et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Huerta Lwanga et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and aquatic environment (Allen et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; C\u0026oacute;zar et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Kibria et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Mart\u0026iacute; et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Indeed, because of the chemical polymers used to manufacture plastic originate from non-biodegradable sources (Andree\u0026szlig;en and Steinb\u0026uuml;chel \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), micrometer-sized particles known as microplastics (MPs) (K\u0026uuml;hnel et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) remain in the environment and are easily absorbed by living organisms, such as humans, plants and fish (S. Singh and Tiwari \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e\u003cp\u003ePlastics can be classified as thermosets and thermoplastics. Thermosets include phenol-formaldehyde resin (bakelite), epoxy (araldite), and polyurethane (PUR), while thermoplastics include polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyethylene terephthalate (PET), polystyrene (PS), high-density polyethylene (HDPE), low-density polyethylene (LDPE), polycarbonate (PC), polyamide (PA), polymethyl methacrylate (PMMA), acrylonitrile butadiene styrene (ABS), acrylonitrile styrene acrylate (ASA), styrene-acrylonitrile copolymer (SAN), and thermoplastic elastomers (TPE) (Plastic Europe n.d.). Thermoplastics are the most prevalent in the environment, as they are the most widely used in the market due to their extensive use in disposable packaging (Cruz and Zanin \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2003\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAs a polymer, plastic degradation processes drastically reduce its molecular mass (C\u0026oacute;zar et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), leading to fissure formation and increased wettability and crystallinity (Waldman and Rillig \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These processes can be physicochemical, biological, and mainly mechanical, due to erosion caused by wind, waves, and solar radiation (photolysis) (Moore \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2008\u003c/span\u003e), generating microplastics (MPs) that are omnipresent and persistent anthropogenic contaminants found in urban environments (Dris et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Furthermore, MPs can cause various pathological alterations due to tissue abrasion by plastic debris, thereby triggering inflammatory responses, insufficiency and reduced energy levels, as well as lower growth rates (Huerta Lwanga et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Y. Song et al. \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Welden and Cowie \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eVarious techniques have been employed to identify MPs based on their chemical composition and specific functional groups (Bolan et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These analyses are conducted using advanced equipment such as stereoscopes, scanning electron microscopes, fluorescence microscopes, Raman/FTIR microscopes, and matrix-assisted laser desorption/ionization (Huppertsberg and Knepper \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Mukotaka et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Siegel et al. \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Y. K. Song et al. \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Fourier-transform infrared (FT-IR) imaging spectroscopy has remained the most widely applied method in microplastic research (Renner et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This method has proven effective for detecting, measuring, and analyzing MP contamination in aquatic and terrestrial communities (Horton et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Napper et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Peeken et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Weisser et al. \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Furthermore, the use of histopathological examinations is necessary to localize microplastics within tissues enhancing the knowledge of its effect in the organism, contrasting with extraction-based approaches that only provide a general overview of particle presence in an organ or tissue (Prata et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). This technique also allows observation of tissue alterations caused by the presence of microplastics in different organs and systems (Lee et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe white-eared opossum (\u003cem\u003eDidelphis albiventris\u003c/em\u003e) is a common marsupial in the municipality of Campo Grande, Mato Grosso do Sul (MS), Brazil (C\u0026aacute;ceres \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Sanches et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). It is considered as synantropic animal because they can be favored by human presence, \u003cem\u003eDidelphis\u003c/em\u003e spp. can be found in urban parks and gardens, vacant lots, in peri-domicile and in human and animal dwellings. Furthermore, as \u003cem\u003eDidelphis\u003c/em\u003e spp. are omnivorous animals, they feed on a wide variety of items, including fruits, insects, eggs, and small vertebrates (C\u0026aacute;ceres \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Contamination by MPs can occur through various routes, such as ingestion, inhalation, dermal contact, transplacental transfer, and predation of contaminated animals. For many mammal species, ingestion is described as the primary contamination pathway (Meaza et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Each contamination route carries specific risks and potential health consequences for the animals and humans (Rahman et al. \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In this context, the present study aimed to investigate the presence of MPs in the gastrointestinal tract and lungs of \u003cem\u003eD. albiventris\u003c/em\u003e sampled in an urban area of Campo Grande, MS. Furthermore, we evaluated the effects of MPs on those tissues and Body Condition (BC) of sampled animals, providing insights into the influence of MPs in the health of this synantropic species in anthropized areas of Mato Grosso do Sul State, Brazil.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cb\u003eStudy Area and Data Collection\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThis study was carried out from April to December 2023 in six forest fragments in the municipality of Campo Grande, MS [Reserva do Surucu\u0026aacute; (20\u0026deg;24'47\"S 54\u0026deg;36'55\"W), CEA Polon\u0026ecirc;s (20\u0026deg;26'49\"S 54\u0026deg;34'48\"W), Clube Carand\u0026aacute; (20\u0026deg;26'21\"S 54\u0026deg;34'27\"W), Ch\u0026aacute;cara Coqueiral (20\u0026deg;29'34\"S 54\u0026deg;34'53\"W), CEA Imbirussu (20\u0026deg;26'57\"S 54\u0026deg;41'48\"W), and CEA Anhandu\u0026iacute; (20\u0026deg;30'22\"S 54\u0026deg;38'37\"W)]. All fragments were sampled at least three times. We used 20 wired live-trap baited with bacon and revised twice a day. Traps remained set for five consecutive nights in each forest fragment, it were installed on the ground and placed approximately 10 meters apart. Captured females were previously evaluated for pregnancy and released if there were cubs attached in their pouch.\u003c/p\u003e\u003cp\u003eIndividuals were anesthetized intramuscularly with ketamine (20 mg/kg) and xylazine (2 mg/kg). Subsequently, individual and biometric data were collected, including head-body length, body-tail length, weight and sex. Afterwards, animals were euthanized by intravenous administration of potassium chloride (75\u0026ndash;150 mg/kg), following the UNIFESP euthanasia protocol. During necropsy, lung, ileum, and stomach (preserving contents) were collected. Samples contemplated for MPs examination were weighed in fragments ranging from 0.13 g to 7.49 g (lung), 0.37 g to 5.76 g (ileum), and 0.5 g to 12.5 g (stomach), wrapped in aluminum foil, and frozen at \u0026minus;\u0026thinsp;20\u0026deg;C until processing. Fragments of the same organs measuring approximately two centimeters in thickness, length, and width were fixed in 10% formalin for histopathological examination.\u003c/p\u003e\u003cp\u003eAll procedures carried out during this research complied with the Biodiversity Authorization and Information System (SISBIO) protocol numbers 70946-5 and 89586-1. Sample handling and processing followed the guidelines established by the Ethics Committee on Animal Use (CEUA) of Dom Bosco Catholic University, protocol number 013/2020.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSample Processing\u003c/b\u003e\u003c/p\u003e\u003cp\u003eStomach, ileum, and lung samples were thawed at room temperature. We used two control filters for each group of organ fragments: the first to monitor air and environmental contamination, and the second was subjected to filtration with 10% KOH solution and Milli-Q water to ensure solution purity. For MPs assessment, biological samples were digested using a 10% potassium hydroxide (KOH) solution (K\u0026uuml;hn et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Samples were rinsed with Milli-Q water to remove residues, weighted on a semi-analytical balance (Marte Ad500 510 g \u0026times; 0.001 g), and placed in glass beakers with 10% KOH solution at a ratio of three times the biological mass (3:1) (Dehaut et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Beakers were covered with aluminum foil and agitated twice daily at room temperature until complete digestion. After digestion, the solution was vacuum filtered using a 47 mm glass fiber microfilter GF-1 (Macherey Nagel) with a pore size of 0.7 \u0026micro;m. Each microfilter was placed individually in glass Petri dishes and dried in an oven for 1 h at 70\u0026deg;C. Filters were examined using a Carl Zeiss optical microscope (MOC), model Axio Scope A1, and images were captured with Zen software (software version to be verified).\u003c/p\u003e\u003cp\u003eTissue images were taken with a color camera (Axiocam 503) attached to the MOC. The materials found were counted and classified following the protocol used by (Clere et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Fibers were measured for length and width, while fragments were measured by area. After filter analysis, filters were washed with Milli-Q water in beakers and pooling samples of all animals were separated by organ. Filters were then oven-dried at 70\u0026deg;C until completely dry. Samples were sent to the Chemistry Institute at the Federal University of Mato Grosso do Sul (UFMS) for FT-IR spectroscopic analysis.\u003c/p\u003e\u003cp\u003e\u003cb\u003eFourier Transform Infrared Spectroscopy\u003c/b\u003e\u003c/p\u003e\u003cp\u003eSpectral data in the mid-infrared region were obtained using a Perkin Elmer Frontier FT/IR spectrometer with Fourier transform. Analyses were acquired over the spectral window from 500 to 4000 cm⁻\u0026sup1;. Samples were prepared by dispersion in KBr pellets. The obtained spectra were compared with those from the reference library created by (Shimadzu n.d.). Only spectra with a match of 70% or higher to the standard database were considered reliable and recorded as verified MPs (Zhao et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2018\u003c/span\u003e)\u003c/p\u003e\u003cp\u003e\u003cb\u003eHistopathological Examination\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFor histopathological examination, fragments of lung, ileum, and stomach were collected. Tissues were sectioned into fragments measuring 3 mm to 5 mm, placed in cassettes, and sent to the Technical Histopathology Center of Curitiba - PR, Brazil, for slide preparation and Hematoxylin and Eosin (H\u0026amp;E) staining. Histopathological slides were analyzed in the laboratory of Dom Bosco Catholic University using a Carl Zeiss optical light microscope (MOC), model Axio Scope A1, and images were captured with the aid of Zen software. Tissue images were photographed using an Axiocam 503 color camera attached to the MOC.\u003c/p\u003e\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStatistical Analysis\u003c/h2\u003e\u003cp\u003eA set of models was constructed to evaluate the effects of MPs size, found in organs analyzed individually and combined, on the body condition (BC) of the collected \u003cem\u003eD. albiventris\u003c/em\u003e. BC was used as a health assessment parameter and calculated by means of the standardized residuals from a linear regression between body weight and head-body length, computed separately for males and females. Additionally, a null model without explanatory variables was created. Generalized linear models (GLMs) were ranked using an approach based on Akaike\u0026rsquo;s Information Criterion corrected for small sample sizes (AICc) (Akaike \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1974\u003c/span\u003e), considering all models with ΔAICc\u0026thinsp;\u0026le;\u0026thinsp;2 as plausible (Burnham and Anderson \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). The \u0026lsquo;AICcmodavg\u0026rsquo; package version 2.3\u0026ndash;1 in R 3.5.0 (R Core Team \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) was used for model ranking.\u003c/p\u003e\u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eOur results show that terrestrial wildlife found in urban areas can be found contaminated by MPs since we found by tissue digestion a total of 270 MPs in the gastrointestinal tract and lungs of the all 22 white-eared opossums captured in the municipality of Campo Grande-MS. In fact, until now, contamination by MPs in terrestrial animals was reported only in domestic animals as sheep (Beriot et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), dogs and cats (Zhang et al. \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), and chicken (Lwanga et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), as well as in earthworms (Lwanga et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Most reports concern MPs contamination refer to marine animals such as bivalves (Li et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), seals (Perez-Venegas et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), commercial fish (Neves et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), pelagic and demersal fish (Neves et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), sea anemones (Morais et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), rays (Pegado et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), and shrimp (Hossain et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Furthermore, since it has been suggested that the accumulation of MPs in the gastrointestinal tract of raptor birds may be due to a combination of direct ingestion of plastics and/or indirect consumption through trophic transfer (Carlin et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). \u003cem\u003eDidelphis albiventris\u003c/em\u003e can also present both routes of contamination by MPs because they feed on small vertebrates and invertebrates. The high number of MPs found in \u003cem\u003eD. albiventris\u003c/em\u003e found in this study may be associated to the omnivorous feeding behavior of this synanthropic species, in accordance to recorded in other vertebrates\u0026rsquo; animals with omnivorous diets, which tend to be broader and less selective (Mazariegos Ort\u0026iacute;z et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Mizraji et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The use of plastic debris from human consumption, such as plastic bags, has been already reported in nests of \u003cem\u003eD. albiventris\u003c/em\u003e (Blettler and Mitchell \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe ileum was the tissue with the highest quantity of microplastics, with 141 MPs recorded (52%), followed by the stomach with 86 MPs (32%) and the lungs with 43 MPs (16%). The high number of MPs in the gastrointestinal tract of \u003cem\u003eD. albiventris\u003c/em\u003e in this study may indicate that ingestion was the most common contamination route. Indeed, for various mammal species, ingestion is considered the primary exposure pathway (Meaza et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), with MPs most frequently detected in the digestive tract, especially in the ileum probably due to the presence of microfold cells located in the epithelium covering mucosa-associated lymphoid tissues, such as the Peyer's patches located in the ileum, that actively transport luminal antigens to the underlying lymphoid follicles to initiate an immune response (Arumugasaamy et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Smith et al. \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e1995\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eFibers were the most common type of MPs found (68.5%), followed by fragments (30.4%), foam (0.7%), and film (0.4%). Fibers was found in 79.4% in ileum sampled, 62.8% lungs, and 53.5% stomach. Studies report a higher prevalence of MPs fibers in the intestinal tract of cows and sheep in Iran (Bahrani et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), intestinal tract of ducks in Indonesia (Susanti et al. \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), as well as in the gastrointestinal tract of terrestrial birds in Florida, USA (Carlin et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) and Shanghai, China (Zhao et al. \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Fibers have high bioaccumulation potential and are among the most common types observed in both marine environments (Browne et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) and soil (R. P. Singh et al. \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). A study on the mussel \u003cem\u003eMytilus galloprovincialis\u003c/em\u003e showed that fibers persisted in the organism longer than fragments and spheres (Park et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Research on zebrafish demonstrated that fiber accumulation in the intestine was greater compared to fragments and spheres, causing more severe intestinal toxicity (Qiao et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Additionally, fibers have been reported in high amounts in lung tissue of pigs and humans with lung cancer (Pauly et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eMP fibers were found in various colors (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), blue was the most abundant one comprising 52% of the particles, appearing most frequently in the lungs and ileum. Transparent MPs were the second most abundant polymer, accounting for 16%, followed by black (14%), amber (7.4%), red (6%), orange (1.8%), purple, and gray (0.7%). Blue fibers have been frequently reported (Araujo Dutra and Camargo Maia \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Mart\u0026iacute; et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) probably because blue plastics tend to degrade faster under sunlight, as they do not absorb UV light effectively, increasing the fragmentation rate and consequently the amount in the environment (Mart\u0026iacute; et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Yang et al. \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe second most common morphological type of MPs was fragments. Like fibers, they were also detected in the lungs, ileum, and stomach. The predominant colors were blue, black, amber, purple, multicolored, transparent, and gray (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The presence of amber coloration in MPs may be related to the photoaging of plastics, which gradually alters their color (discoloration) during prolonged solar exposure (Mart\u0026iacute; et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), considering that detection of amber-colored MPs in the gastrointestinal tract of animals is rarely reported. Plastic color significantly affects the absorption of sunlight, with plastics of different colors absorbing light at different wavelengths (Zhao et al. \u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eRegarding FT-IR analyses, three types of polymers were identified: polyethylene terephthalate (PET), acrylonitrile butadiene styrene (ABS), and polyvinyl chloride (PVC). These polymers are classified as thermoplastics, the most commonly used in disposable packaging industries (Plastic Europe n.d.). We observed that all samples contained PVC (Supplementary Material \u0026ndash; Figures \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e, S2 and S3). While PET was found in the ileum and lungs (Supplementary Material \u0026ndash; Figures S2 and S3), ABS was detected only in lungs (Supplementary Material \u0026ndash; Figure S3).\u003c/p\u003e\u003cp\u003eThe presence of PVC and PET in the gastrointestinal tract and lungs of \u003cem\u003eD. albiventris\u003c/em\u003e can be attributed to the high abundance of these polymers in the environment (Akdogan and Guven \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), as they are among the most commonly found MPs in both soil (Yang et al. \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) and air (Yao et al. 2022). Indeed, \u003cem\u003eD. albiventris\u003c/em\u003e are frequently found in trash bins in Campo Grande, potentially ingesting PVC and PET from plastic commonly used packaging (Wang et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The presence of ABS in animals has not been previously reported in the literature. Furthermore, reports of this polymer in sediments and effluents, are also scarce (Belesov et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Sarti et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Since we found ABS only in the lungs, it seems that the airborne contamination route would be the most likely.\u003c/p\u003e\u003cp\u003eWe highlight that MPs have been identified in a wide range of species (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), from urban dogs and cats to dolphins, birds of prey, terrestrial mammals, and commercial fish, covering different geographic regions and habitats. The increasing detection of microplastics in various tissues and organs of animals reinforces the urgency of studies focused on pollution caused by these emerging contaminants. The most common polymers include PET, PE, PP, and PVC, with a predominance of synthetic fibers and fragments smaller than 1 mm. The presence of these materials in vital organs such as lungs, liver, gastrointestinal tract, and reproductive tissues raises concerns about their toxic, bioaccumulative effects and potential consequences on animal health and, by extension, human health, given the ecological interconnectedness. These data emphasize that the ubiquity of MPs in the environment is already directly reflected in biota, making it essential to deepen investigations into exposure mechanisms, accumulation, and toxicity, as well as to strengthen public policies and mitigation actions against this form of pollution.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eStudies on the detection of microplastics in tissues and organs of different animal species across various countries.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAnimals\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLocation\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePolymer Types\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eOrgans\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eTechniques Uses\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eRemarks\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eAuthors\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWhite-eared opossum\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMato Grosso do Sul, Brazil\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePET, PVC and ABS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eStomach, ileum, lung\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFT-IR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e100% showed MPs in at least one organ, highlighting their role as sentinels of contamination, potentially exposed via ingestion and inhalation.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eThis study\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eUrban dogs and cats\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePorto, Portugal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePET, PP\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLung, ileum, liver, kidneys, blood clots\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eMicro-Raman spectroscopy\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eMicroplastics were found in 100% of the internal tissues analyzed, predominantly synthetic fibers, indicating high environmental exposure and potential health risks to urban animals.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003ePrata et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2022\u003c/span\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDogs and humans\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNew M\u0026eacute;xico, USA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePE, PVC, PET\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eTesticular tissues\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePy-GC/MS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003ePresence of MPs in the testes, decreased sperm count, and reduced testicular and epididymal weight, suggesting potential impacts on male reproductive health\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eHu et al., 2024\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTerrestrial birds of prey\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFlorida, USA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePET, PE\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGI tract\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u0026micro;-FTIR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eGeneralized accumulation of MPs in the gastrointestinal tract, with 100% of samples contaminated, indicating wide environmental exposure and potential health risks.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eCarlin et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWild terrestrial animals\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eThailand\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePE, PET, PP, PVDC, PES, PU\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eCarcasses (organs not specified)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFT-IR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eMPs found in 71% of fecal samples, demonstrating that plastic contamination already widely affects the terrestrial fauna in the region.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eTeampanpong \u0026amp; Duengkae, 2024\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDolphins (\u003cem\u003eTursiops truncatus\u003c/em\u003e)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFlorida, USA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePolyester\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGastrointestinal fluid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFT-IR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003ePresence of MPs in muscular and gastrointestinal tissues of fish from Sarasota Bay, the dolphins' main food source, suggesting risk of indirect exposure.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eHart et al., 2022\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCommercial fish\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eXisha, China\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePE, PET, PVC, PP, PVA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGills and GI tract\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFT-IR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eMPs detected in 61.7% of reef fish, primarily in the gastrointestinal tract, with a predominance of polyamide and PET fibers.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eHuang et al., 2023\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWild coastal mammals, birds, and fish\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBergen, Norway\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePET, PVC, PC, PP, PE, PS, PMMA, PA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eStomach, intestinal wall, liver, muscle\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePy-GC/MS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eMPs present in 62% of analyzed tissues, with higher concentrations in the liver and a predominance of PVC, PS, and PET, indicating potential bioaccumulation in internal organs.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eHaave et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCoyote, Virginia opossum, white-nosed coati, white-tailed deer, Mexican and lesser anteater\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eYucat\u0026aacute;n, Mexico\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eElastomers, PS, HDPE, cellulosic polymers, PU\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFeces\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFT-IR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eMPs detected in 100% of terrestrial vertebrate fecal samples, predominantly fibers\u0026thinsp;\u0026lt;\u0026thinsp;1 mm composed of polyamide and PET, revealing widespread environmental contamination.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMendoza-Arroyo et al., 2024\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTerrestrial mammals\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eUnited Kingdom\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePolyester, PE, PNB, and biodegradable plastics\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFeces\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u0026micro;-FTIR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eThe study revealed that 16.5% of the terrestrial mammals ingested MPs, highlighting extensive exposure to environmental pollution.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eThrift et al., 2022\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eNote.\u003c/b\u003e PET\u0026thinsp;=\u0026thinsp;polyethylene terephthalate; PE\u0026thinsp;=\u0026thinsp;polyethylene; PP\u0026thinsp;=\u0026thinsp;polypropylene; PVC\u0026thinsp;=\u0026thinsp;polyvinyl chloride; PS\u0026thinsp;=\u0026thinsp;polystyrene; PU\u0026thinsp;=\u0026thinsp;polyurethane; PVA\u0026thinsp;=\u0026thinsp;polyvinyl acetate; PMMA\u0026thinsp;=\u0026thinsp;polymethyl methacrylate; PNB\u0026thinsp;=\u0026thinsp;polybutylene; ABS\u0026thinsp;=\u0026thinsp;acrylonitrile butadiene styrene. FT-IR\u0026thinsp;=\u0026thinsp;Fourier-transform infrared spectroscopy; \u0026micro;-FTIR\u0026thinsp;=\u0026thinsp;micro-Fourier-transform infrared spectroscopy; Py-GC/MS\u0026thinsp;=\u0026thinsp;pyrolysis coupled with gas chromatography and mass spectrometry. GI tract\u0026thinsp;=\u0026thinsp;gastrointestinal tract.\u003c/p\u003e\u003cp\u003eThe histopathological examination of sampled \u003cem\u003eD. albiventris\u003c/em\u003e revealed the presence of few MPs in only intestinal interstitium and the lumen of the stomach, amid the contents, with no inflammatory reaction observed in the organs (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). These findings are in agreement with Haave et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) that did not find MPs or tissue reaction by histopathological examination in fish, seabirds, terrestrial and marine mammals from a coastal area of Norway. The number of MPs observed in the histopathology of the gastrointestinal tract of \u003cem\u003eD. albiventris\u003c/em\u003e, in this study, seems insufficient to cause intestinal blockage or a feeling of fullness. The pulmonary parenchyma of \u003cem\u003eD. albiventris\u003c/em\u003e did not showed neither MPs nor alterations. Most studies evaluating histopathological effects of MPs are experimental, conducted in laboratory settings (Ali et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Hamed et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), aimed to induce tissue damage (Hoseini et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Saleh et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Sayed et al. \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These studies use high concentrations of commercial MPs, that are not commonly found in the environment (Allen et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Sun et al. \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe contamination of MPs in \u003cem\u003eD. albiventris\u003c/em\u003e does not seem to threaten health of sampled animals since our statistical analyses did not correlate the presence of MPs in tissues with BC (Supplementary Material, Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). This corroborates with de Vries et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) in cod and hake contaminated with MPs, which hypothesized that, especially in larger individuals, MPs are not retained, thus not affecting BC, unlike what occurs in small fish species (Critchell and Hoogenboom \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eAll individuals of \u003cem\u003eD. albiventris\u003c/em\u003e were found contaminated by MPs, without effects on health of sampled animals. Moreover, this synanthropic species may be used as sentinels of environmental contamination in anthropized areas.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eEthical Approval\u003c/h2\u003e\u003cp\u003e The research was approved by the Animal Use Ethics Committee of Dom Bosco Catholic University (protocol 013/2020) and the Biodiversity Authorization and Information System (SISBIO) (protocol 70946-5 and 89586-1). The authors declare no relevant financial or non-financial interests.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConflict of Interest Statement\u003c/strong\u003e\u003cp\u003eThe author(s) declared no potential conflicts of interest regarding the research, authorship, and/or publication of this article.\u003c/p\u003e\u003cp\u003e\u003ch2\u003eAuthors\u0026rsquo; Ethical Responsibilities\u003c/h2\u003e\u003cp\u003e All authors have read, understood, and complied, as applicable, with the statement on \"Authors' Ethical Responsibilities,\" as found in the Instructions for Authors.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding Declaration\u003c/h2\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThis work was supported by National Council for Scientific and Technological Development (CNPq) and Coordination of Improvement of Higher Education Personnel (CAPES). ACR received a fellowship from CNPq (163253/2022-06). FMS received a fellowship from Funding Authority for Studies and Projects (FINEP) (01.24.0114.00) and CNPq (Productivity Scholarship 302176/2025-0), PSC received a fellowship from Ministry of Science, Technology and Innovation (MCTI) (352739/2025-8) and HMH is researcher from CNPq (Productivity Scholarship 311769/2023-3).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eF.M.G.M., A.C.R., G.B.A., P.S.C., and H.M.H. contributed to the conceptualization of the study. F.M.G.M., P.S.C., A.C.R., O.F.J., and W.A.G.N. were responsible for the methodology. P.S.C., A.C.R., G.B.A., F.M.S., and T.G.S.P. conducted formal analysis and investigation. F.M.G.M., P.S.C., H.M.H., G.B.A., N.Y.S., and F.M.S. contributed to writing, review and editing. H.M.H. and P.S.C. provided resources. G.B.A., H.M.H., and P.S.C. supervised the project. All authors reviewed the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e\u003cp\u003eWe thank the support provided by Interface Research Group Between Human, Animal and Environmental Health research group (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ewww.insanahuna.com\u003c/a\u003e\u003c/span\u003e\u003cspan address=\"http://www.insanahuna.com\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe data will be made available on request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAkaike, H. (1974). A New Look at the Statistical Model Identification. 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Microplastics in soils: A review of methods, occurrence, fate, transport, ecological and environmental risks. \u003cem\u003eScience of The Total Environment\u003c/em\u003e, \u003cem\u003e748\u003c/em\u003e, 141368. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scitotenv.2020.141368\u003c/span\u003e\u003cspan address=\"10.1016/j.scitotenv.2020.141368\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"environmental-monitoring-and-assessment","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"emas","sideBox":"Learn more about [Environmental Monitoring and Assessment](http://link.springer.com/journal/10661)","snPcode":"10661","submissionUrl":"https://submission.nature.com/new-submission/10661/3","title":"Environmental Monitoring and Assessment","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Bioindicator, Body condition, Emerging contaminant, Histopathology, Polyethylene, Wild mammal","lastPublishedDoi":"10.21203/rs.3.rs-7284106/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7284106/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study aimed to investigate the presence of microplastics in the gastrointestinal tract and lungs of white-eared opossums (\u003cem\u003eDidelphis albiventris\u003c/em\u003e), as well as their effects on these organs and body condition of sampled animals. We captured 22 opossums from April to December 2023 in six forest fragments of Campo Grande, capital of Mato Grosso do Sul state. The estimate of body condition was performed using measurements of weight and head-body length for males and females. Stomach, ileum, and lung fragments were collected during necropsies for histopathological study and detection of microplastics by alkaline digestion with KOH. Digested samples were vacuum filtered using filter paper and dried in an oven. The dried filters were examined under an optical microscope. A total of 270 microplastics were detected in the 22 animals examined. The ileum presented the highest number of microplastics (n\u0026thinsp;=\u0026thinsp;141), fibers were the most common type (n\u0026thinsp;=\u0026thinsp;185), and blue was the predominant color (n\u0026thinsp;=\u0026thinsp;140). Regarding chemical composition, the most frequently detected microplastics were polyvinyl chloride, polyethylene terephthalate, and acrylonitrile butadiene styrene. Although all \u003cem\u003eD. albiventris\u003c/em\u003e presented microplastics in their evaluable tissues, no effects on body condition or tissue damage were observed. Since \u003cem\u003eD. albiventris\u003c/em\u003e are common animals in Campo Grande and are contaminated by microplastics, these animals can act as sentinels of environmental condition.\u003c/p\u003e","manuscriptTitle":"Contamination by Microplastics in Free-Living White-Eared Opossums (Didelphis Albiventris) Resident in Campo Grande, Mato Grosso Do Sul – Brazil","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-22 12:23:48","doi":"10.21203/rs.3.rs-7284106/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-09T10:47:42+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-08T23:37:41+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-26T20:23:58+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-22T21:05:37+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"42251511863119357668282630499247621669","date":"2025-08-18T19:05:08+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"305006846117639906260607072478544937673","date":"2025-08-18T15:02:55+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"54840476558659836835741628920377927236","date":"2025-08-15T16:37:38+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-08-15T10:30:04+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-08-08T23:08:18+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-08-08T23:07:45+00:00","index":"","fulltext":""},{"type":"submitted","content":"Environmental Monitoring and Assessment","date":"2025-08-03T14:44:43+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"environmental-monitoring-and-assessment","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"emas","sideBox":"Learn more about [Environmental Monitoring and Assessment](http://link.springer.com/journal/10661)","snPcode":"10661","submissionUrl":"https://submission.nature.com/new-submission/10661/3","title":"Environmental Monitoring and Assessment","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"6eac069c-a431-486b-981c-cbf172fa69de","owner":[],"postedDate":"August 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-10-27T16:33:03+00:00","versionOfRecord":{"articleIdentity":"rs-7284106","link":"https://doi.org/10.1007/s10661-025-14682-3","journal":{"identity":"environmental-monitoring-and-assessment","isVorOnly":false,"title":"Environmental Monitoring and Assessment"},"publishedOn":"2025-10-22 16:17:10","publishedOnDateReadable":"October 22nd, 2025"},"versionCreatedAt":"2025-08-22 12:23:48","video":"","vorDoi":"10.1007/s10661-025-14682-3","vorDoiUrl":"https://doi.org/10.1007/s10661-025-14682-3","workflowStages":[]},"version":"v1","identity":"rs-7284106","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7284106","identity":"rs-7284106","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}